New 1100 Series DAD SL and MWD SL 80Hz Data Acquisition for Ultra-fast LC

Series DAD SL and MWD SL Agilent 1100 Series Diode-array Detector SL Agilent 1100 Series Multi-wavelength Detector SL The 1 st Diode-array Detector designed for Ultra-fast LC

Series DAD SL and MWD SL Overview Next Generation Electronics and Firmware provides  80Hz Data Acquisition of up to 8 Signals Up to 100% resolution gain in ultra-fast, quantitative LC  80Hz Full Spectral Data Acquisition (DAD SL only) for ultra-fast peak purity analysis and spectral conformation even for trace level compounds  Improved Diode-Array Front-end Electronics for minimized noise (typical < +/- 6μAU ASTM)  New Build-in Data Recovery Card for a „data never lost insurance“  New RFID Tags for all Flow Cells and UV Lamp for unambiguous data traceability  LAN on Board eliminates need for additional LAN interface  Future proof design Build-in web-server, USB, PCMCIA (WLAN, Bluetooth)

Series DAD SL and MWD SL Overview (cont’d)  New Electronic Temperature Control (ETC) For maximum practical sensitivity by minimized baseline wander, especially under harsh and fluctuating ambient temperature and humidity conditions  New standard flow cell for maximum practical sensitivity by minimized RI-sensitivity and dispersion  Builds upon 1100 DAD Optical Design preserves features like programmable slit and dual lamp design for highest sensitivity from 190 to 950nm  Up to 20x Sensitivity from 400 – 950nm  + 40% (+ 80%) Sensitivity by 8 and 16nm slit Re-use spectral libraries of 1100 DAD „A“ and „B“ Confidence and robustness of a diode-array detector with more than 25,000 installations.

5 What is Ultra-fast LC? What is the Objective of Ultra-fast LC? 1. Significant gains in productivity, while maintaining or increasing data quality Ultra-fast LC provides up to 10x gains in analysis speed while preserving or increasing Resolution, Sensitivity, Linearity, Precision and Robustness. Thereby, Ultra-fast LC ensures compliance with strictest (regulatory) performance requirements. 2. Uncompromised compatibility with existing methods. Run conventional LC methods without compromising performance Comply with today’s and future requirements 3. Improved Data Security. Ultra-fast LC systems provide a new level of data security and traceability that prevents data losses and minimizes the risk of false data interpretation.

6 Conventional LC: Analysis Times =5.0 – 120 min Fast LC: Analysis Times= 2.0 – 5.0 min Ultra-fast LC:Analysis Times =0.2– 2.0 min Gradient Time= 0.2– 1.5 min Cycle Times =0.5– 2.5 min 50% Peak Width=0.1– 1.0 sec PW = 0.3 sec min How fast is Ultra-fast? What is Ultra-fast LC?

7 Which 1100 Configuration do I need for Ultra-fast LC? 1100 Series Ultra-fast LC System 1100 Series Binary Pump – for precise, high-pressure mixing gradient formation and low delay volume 1100 Series WPS – for precise, high-speed injection with lowest delay volume and carry over 1100 Series TCC – for precise, peltier-controlled high-temperature LC up to 80C 1100 Series DAD/MWD SL – for highest chromatographic resolution by 80Hz data rate Zorbax RRHT 1.8um Columns – for highest efficiency at high linear flows What is Ultra-fast LC?

8 min Hz PW=0.30sec 40Hz PW=0.33sec 20Hz PW=0.42sec 10Hz PW=0.67sec 5Hz PW=1.24sec Sample: Phenone Test Mix Column: Zorbax SB-C18, 4.6x30, 1.8um Gradient: % ACN in 0.3min Flow cell: 5ul 80Hz versus 20Hz – 30% Peak Width + 30% Resolution + 40% Peak Capacity + 70% Apparent Column Efficiency 80Hz versus 10Hz – 55% Peak Width + 90% Resolution + 120% Peak Capacity + 260% Apparent Column Efficiency What’s the Benefit of 80Hz Data Acquisition Rate? Peak Width, Resolution and Peak Capacity in Ultra-Fast LC

9 80Hz versus 20Hz Data Rate: –30% Peak Width=>+40% Peak Capacity +30% Resolution =>+ 70% Apparent Column Efficiency 80Hz versus 10Hz Data Rate: – 55% Peak Width=>+120% Peak Capacity +90% Resolution =>+260% Apparent Column Efficiency Data Rate Peak Width ResolutionPeak Capacity 80 Hz Hz Hz Hz Hz What’s the Benefit of 80Hz Data Acquisition Rate? Peak Width, Resolution and Peak Capacity in Ultra-Fast LC

10 Performance Requirements of Ultra-Fast LC Ultra-fast LC using the 1100 DAD SL provides Resolution and Peak Capacity gains of up to 100%. But – Can I still fulfill my (regulatory) performance requirements under ultra-fast LC conditions? Quantification of Side Products at 0.05% level ? RT Precision < 0.5% ? Area Precision < 1% ? Peak Purity Analysis at Trace Levels ? Spectral Conformation at Trace Levels ?

11 Sensitivity and Linearity in Ultra-Fast LC Can I Simultaneously Quantity Main Compounds and Side Products at 0.05% Level? min mAU Main Compound = 2000mAU Impurities = 1mAU DMSO Impurities Is the detectors Dynamic Range large enough to accurately and precisely quantify Main Compound and Impurities simultaneously?

12 Sensitivity and Linearity in Ultra-Fast LC Is the Noise Low Enough for my Quantitative Analysis? ASTM Noise Specification: 20 µAU Peak-to-Peak (+/- 10 µAU) 4nm Slit8nm Slit16nm Slit 80 Hz Hz Hz Hz Hz Temperature: 20C DAD: 254nm,16nm, Ref 360, 80nm PW: > 0.1min (2.5Hz, 2sec RT) min mAU min mAU Conditions Eluent: Water/ACN = 70/30 Flow rate: 1ml/min Column: 4.6x30mm SB C18, 1.8um min mAU nm Slit width Noise < +/– 3.7 µAU 8nm Slit width Noise < +/– 2.7 µAU 16nm Slit width Noise < +/– 2.0 µAU Peak-to-Peak Noise on 13ul Flow Cell – 27% – 26% Note:50µAU Noise gives S/N = 20 at 1mAU (0.05% level)

13 Sensitivity and Linearity in Ultra-Fast LC Is the Linear Range Large Enough for my Quantitative Analysis? Linearity (Caffeine Sample): Deviation at 2.0AU: 2.0% (Vis Lamp off) 2.5% (Vis Lamp on) 5% Deviation: 2.5 AU (Vis Lamp off) 2.4 AU (Vis Lamp off) Specification: 5% Deviation at 2.0 AU

14 Gradient: 50–70% B in 0.85min Column: 4.6 x 50, 1.8um Injection: 5ul of 550 µg/ml Nimodipin Flow Rate: 4 ml/min Flow cell: 13ul Data Rate: 80Hz Slit: 8nm Overlay of 10 analyses at 245nm RT Precision: 0.067% RSD Area Precision: 0.13% RSD Sensitivity and Linearity in Ultra-Fast LC Reproducibility of Main Compounds at 2000mAU (100% Level)

15 Sensitivity and Linearity in Ultra-Fast LC Reproducibility of Impurities and Side Products at Trace Level Overlay of 10 analyses at 245 nm: Nifedipin A = 2.5mAU (0.1% level) RT Precision = 0.092% RSD Nifedipin degradation product A = 0.5mAU (0.03% level) RT Precision = 0.123% RSD Column: 4.6 x 50, 1.8um Gradient: 50–70% B in 0.85 min Injection: 5ul Flow Rate: 4 ml/min Flow cell: 13ul Data Rate: 80Hz Slit: 8nm

16 Nifedipin at trace levels Peak Width = 0.63 sec min mAU mAU = 0.1% level S/N = 50 1mAU = 0.05% level S/N = mAU = 0.025% level S/N = 12 Conditions: Column: 4.6 x 50, 1.8um Gradient: 50–70% B in 0.85 min Injection: 5ul Flow Rate: 4 ml/min Flow cell: 13ul Data Rate: 80Hz Slit: 8nm Result: Under ultra-fast LC conditions the DAD SL allows accurate quantitation of impurities and side products at levels smaller than 0.05% of the main compound(s). Sensitivity and Linearity in Ultra-Fast LC Can I quantity Impurities and Side Products at 0.05% Level? Noise 40 µAU

17 Apex Spectrum of Nifedipin at 0.1% (1.8 mAU) measured at 80 Hz: nm mAU nm mAU Reference Spectrum of Nifedipin at 100% level (1800 mAU) measured at 80 Hz: Spectral Analysis in Ultra-Fast LC Can I do Peak Purity and Spectral Conformation at Trace Levels? Is the spectral quality obtained at trace level under ultra-fast LC conditions and 80Hz spectral sampling rate good enough for peak purity analysis and spectral conformation?

18 UV Spectrum of Nifedipin at higher concentration (ca. x180): Spectral Analysis in Ultra-Fast LC Can I do 80Hz Peak Purity Analysis at Trace Levels? mAU Overlay of extracted Nifedipin spectra at trace level Result: Nifedipin peak at 0.1% level (1.8mAU) measured with 80Hz spectral rate is pure – no other compounds are co-eluting with Nifedipin

19 nm Norm nm Norm Overlay of 80Hz Reference and Apex Spectrum Nifedipin Ref: 1800mAU (100% level) Nifedipin Apex: 1.8mAU (0.1% level) Spectral Analysis in Ultra-Fast LC Can I do 80Hz Spectral Conformation at Trace Levels? Overlay of 80Hz Reference and Apex Spectrum Nimodipin Ref: 1800mAU (100% level) Nifedipin Apex: 1.8mAU (0.1% level) Result: Identification of Nifedipin at 0.1% trace level under fast LC conditions and 80Hz spectral sampling rate Match Factor = 963Match Factor = 929

20 Performance Average 50% Peakwidth = 0.34 sec Resolution (4,5) = 1.5 Analysis Time = 24 sec Cycle time = 50 sec RT Precision = 0.7 – 0.22% RSD Area Precision = 1.5 – 0.3% RSD Conditions – An extreme Example Sample: Phenone Test Mix Column: 4.6 x 30mm, 3.5µm SB-C18 Gradient: % ACN in 0.3min Flow rate: 5ml/min Temperature: 40°C Data Rate: 40Hz Speed and Precision in Ultra-Fast LC Pushing the Limits for highest Throughput PeakRSD RT (%)RSD Area (%) Overlay of 6 Runs Application Areas Screening Experiments HT LC/MS/UV Early Formulation Studies Process Analytical Techn.

21 Speed and Precision in Ultra-Fast LC Moderate Gradients for highest-quality Quantitative Data PeakRSD RT (%)RSD Area (%) min mAU Overlay of 6 Runs 2x slower gradient 2 – 5x better Precision Still very fast Performance Peakwidth = 0.38 sec FWHM Resolution (4,5) = 1.6 Analysis Time = 40 sec Cycle time = 70 sec RT Precision = 0.1 – 0.5% RSD Area Precision = 0.2 – 0.5% RSD Conditions – Less extreme Condtions Sample: Phenone Test Mix Column: 4.6 x 30mm, 3.5µm SB-C18 Gradient: % ACN in 0.6min Flow rate: 5ml/min Temperature: 40°C Data Rate: 40Hz Application Areas Formulation Studies Analytical Development Process Control QA/QC

22 min injection 1 injection 4000 Column 2 Column 1 injection 4000 injection 2000 injection 1 min Robustness in Ultra-Fast LC Are Methods and Instrumentation Robust Enough for 24x7 Operation? Stability study on system configuration with automated column regeneration: Stable system and column performance for 8000 injection (4000 injections per column) System suitable for unattended and automated 24x7 operation and reliable over-weekend runs P=300bar

23 New Data Recovery Card* The first LC Detector with “Data Never Lost” Insurance *Patent Filed Data Recovery Card* - DRC All signals, spectra and meta data are buffered on high-capacity, embedded 256MB Compact Flash Card compliant with 21 CFR Part 11. Prevents any data loss in case of communication breakdowns between instrument and PC. Automatic Run Recovery in case of temporary communication failures Manual Run Recovery in case of permanent communication failures” after software, PC, and/or instrument re-boot.

24 New Data Recovery Card The first LC Detector with “Data Never Lost” Insurance Automated Run Recovery in case of temporary communication failures A) Time Elapsed = 0.3 min Run in Progress B) Time Elapsed = 0.6 min Communication Failure Occurs C) Time Elapsed = 1.9 min Communication is re-established Automatic data transfer from DRC to PC No user interaction necessary

25 New Data Recovery Card The first LC Detector with “Data Never Lost” Insurance Manual Run Recovery in case of permanent communication failures Run Recovery dialog pops-up automatically after system re-boot. Data are stored on the PC under a pre-configured location.

26 The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and UV Lamp Radio Frequency Identification Tags RFID tags records all relevant data necessary to recall instrument conditions under which a run has been executed. Minimizes the risk of false data interpretation, because measurement conditions are documented. Meta data stored on RFID tags are saved with each raw data file for unambiguous answers to (auditor-) questions like “Which type of flow cell was used to generate this chromatogram - what was the path length and volume?” “Did the accumulated burn-time of the lamp exceed pre-defined limit?” Flow Cell Path length Volume Max pressure Date last test passed Product number Serial number Production date UV Lamp Accumulated on-time Actual on-time Number of ignitions Date last test passed Product number Serial number Production date

27 The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and UV Lamp ChemStation Report RFID-tag information documents run conditions.

28 The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and UV Lamp 10 RFID Tag Info for Diagnostics

29 The next Level of Baseline Stability New Electronic Temperature Control – ETC Electronic Temperature Control - ETC Advantages of ETC Compensation of changes of ambient conditions (temperature and humidity) Reduced baseline wander for improved practical sensitivity and reproducibility under harsh environmental conditions Optical UnitMain Board Optical Temp. Sensor Ambient Temp. Sensor Air Flow FanHeater 1100 Series DAD SL / MWD SL Front

30 Conditions: Relative humidity = 60%RH = const; Temp = 25°C +/-2°C; 4 x 1h Cycles Note: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition). TempCntrl ON 254,4 No Ref 750,4 No Ref TempCntrl OFF 254,4 No Ref 750,4 No Ref 2.6mAU / 4°C = 0.7mAU/°C The next Level of Baseline Stability New Electronic Temperature Control – ETC Ambient Rejection at 60% RH: ETC off: ~ 700µAU/°C ETC on: < 30µAU/°C Temp. C RH % AH g/kg AH Dev. %

31 TempCntrl ON 254,4 No Ref 750,4 No Ref TempCntrl OFF 254,4 No Ref 750,4 No Ref ~0.5mAU/°C ~0.7mAU/°C Conditions: Relative humidity = 95% RH = const; Temp = 25°C +/-2°C; 4 x 1h Cycles Note: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition). The next Level of Baseline Stability New Electronic Temperature Control – ETC Ambient Rejection at 95% RH: ETC off: ~ 700µAU/°C ETC on: < 30µAU/°C Temp. C RH % AH g/kg AH Dev. %

DAD SL: 254,4 / No REF 1100 DAD SL: 254,4 / 360, DAD B: 254,4 No REF1100 DAD B: 254,4 / 360,100 ~100µAU/°C Comparison between 1100 DAD SL and 1100 DAD (“B”-model) Conditions: Relative humidity = 60%RH = const; Temp = 25°C +/-2°C; 4 x 1h Cycles Note: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition). The next Level of Baseline Stability New Electronic Temperature Control – ETC Ambient Rejection ETC on: DAD “B”: ~ 100µAU/°C DAD SL: < 30µAU/°C Temp. C RH % AH g/kg AH Dev. %

33 - Insert - Body - Ring New 13ul Standard Flow Cell Design* Ceramic Ring for thermal de-coupling *Patent Filed “Drill” Design of Inlet and Outlet for faster flush-out Advantages Reduced RI-sensitivity Improved peak dispersion Minimized noise in in high-flow, high-temperature applications

34 New 13ul Standard Flow Cell Reduced RI Sensitivity New Standard Flow Cell provides 3-4x lower RI-Sensitivity 1100 DAD/MWD with old Standard Cell 1100 DAD SL/MWD SL with new Standard Cell 12 mAU 4 mAU 36 mAU 9 mAU Demanding RI Test Gradient

35 Column Sample Temp Flow WL Eluent Injection SB-C18 1.0x50 Iso Std +H 2 NCSNH ml/min 254,4; -/- 80/20 H 2 O/ACN 1 ul New 13ul Standard Flow Cell Minimized Dispersion for Maximum Resolution and Sensitivity Improved flush-out behavior of the new standard flow cell minimizes peak tailing thereby increasing peak heights and resolution. new Standard Cell old Standard Cell

36 - Insert - Body - Ring Flow Cell Portfolio of the 1100 DAD/MWD SL For Uncompromised Compatibility from Nano to Prep Advantages – From Nano to Prep Compatibility with Conventional LC, Ultra-fast LC, Capillary LC, Nano LC and Prep LC Support of Analytical LC on Columns from 75 µm to 4.6mm ID Support of Preparative LC on Columns from 4.6 to 50mm ID RFID tags for Data Traceability and Diagnostics Flow Cell Portfolio  Standard: 13ul, 10mm path length, 120bar  Semi-Micro: 5ul, 6mm path length, 120bar  Micro: 1.7ul, 6mm path length, 400bar  Semi-Nano: 500nl, 10mm path length, 50bar  Nano: 80nl, 6mm path length, 50bar  Preparative: 3mm, 120bar  Preparative: 0.3mm, 20bar  Preparative: 0.06mm, 20bar

37 Which Flow Cell to use for Ultra-fast LC? 13µl Standard Flow Cell:  For highest sensitivity and linearity  High-demanding quantitative work, e.g. analytical method development, QA/QC  4.6 – 3 mm ID Columns 1.7µl Micro Flow Cell:  For highest selectivity  Ultra-fast semi-quantitative work, e.g. Screening Experiments, HT LC/MS/UV  2.1 – 1mm ID Columns 5µl Micro Flow Cell:  Best compromise of sensitivity and selectivity  For good quantitative and qualitative results, e.g. Screening, HT LC/MS/UV, Early Formulation Studies  4.6 – 1mm ID Columns Flow Cell Volume/Pathlength Sensitivity & Linearity Resolution 13 µl / 10mm100 %~ 90 – 95% * 5 µl / 6mm~ 75 – 85% *~ 93 – 98% * 1.7 µl / 6mm~ 65 – 75% *100 % * Depends on analytical conditions and column dimension

38 min mAU Which Flow Cell to use for Ultra-fast LC? Noise Comparison 13ul Standard Flow Cell Noise < +/- 2.1 uAU 5ul Semi-Micro Flow Cell Noise < +/- 2.4 uAU Conditions Column: 4.6x30mm SB C18, 1.8um Flow rate: 1ml/min Mobile phase: Water Temperature: 20C DAD: 254nm, 16nm, Ref 360, 80nm PW: > 0.1min (2.5Hz, 2sec RT) Slit = 16 min mAU Specification: ASTM noise < +/- 10 uAU Note: 5ul cell shows similar noise as 13ul cell. However, the linear range is reduced due to the reduced path length of 6mm.

39 Sensitivity Limit of Detection for Anthracene under Ultra-fast LC Conditions 10pg Anthracene injected in 1ul 4nm slit 40Hz: LOD = 1.48pg 20Hz, LOD = 1.03pg 10Hz, LOD = 0.67pg 80Hz: LOD = 2.30pg mAU min Performance: LOD/pg in Ultra-fast LC Retention Time = 12 sec, Peakwidth = 0.9 sec 4nm Slit8nm Slit16nm Slit 80 Hz Hz Hz1.030, Hz Hz – 27% – 26% 4nm Slit8nm Slit16nm Slit 2.5 Hz Compare: Conventional LC on 1100 DAD/MWD „B“ Retention Time = 2 min, Peak width = 6 sec

Series DAD SL and MWD SL – Builds upon 1100 DAD Optical Design - Optimized for Best Sensitivity Tungsten lamp Long-life Deuterium Lamp Holmium Oxide Filter Flow Cell Programmable slit Grating 1024 element diode array 190 nm 950 nm Minimized Noise in Visible WL-Range Automated wavelength verification Fast optimization of sensitivity and resolution Excellent wavelength resolution More uptime > 2000h 1100 DAD – 25,000 Installations Worldwide

41 Tungsten Lamp Off: Noise: > 400 µAU 700nm 254nm ACN/Water 20/ ml/min 1 nm Eluent: Flow rate: Slit: 1100 Series DAD SL and MWD SL – Builds upon 1100 DAD Dual Lamp Design for Highest Sensitivity from 190 to 950nm Tungsten Lamp On Noise: < 20 µAU Tungsten Lamp Off: Noise: < 20 µAU mAU min Tungsten Lamp On Noise: < 20 µAU Advantages Approx. constant Noise from 190 to 950nm Up to 20x lower Limits of Detection for compounds absorbing in the visible range at λ > 400nm Significantly higher confidence in qualitative, spectral analysis results  More accurate Peak Purity results, especially at trace levels.  More accurate Library Analysis and Spectral Confirmation results

Series DAD SL and MWD SL – Builds upon 1100 DAD Programmable Slit – Micromechanics Motor Slit 1 Programmable Slit for 1, 2, 4, 8 and 16 nm Advantages No need for manual slit changes Fast optimization of sensitivity and resolution  Maximized Sensitivity by 16nm slit (+ 80% versus 4nm slit)  Maximized Spectral Resolution by 1nm slit (for optimized spectral analysis) Documents slit width (GLP, data traceability)

43 nm Resolution Benzene Spectrum at Trace Level 0.7 mAU 1 nm slit 2 nm wavelength bunching 1 nm 2 nm 4 nm 8 nm Absorbance (mAU) 16 nm 1100 Series DAD SL and MWD SL – Builds upon 1100 DAD Programmable Slit for fast Optimization of Sensitivity and Resolution Noise level

44 Rapid Resolution HT 1100 Series Modification Kits Converting an 1100 Binary System to Ultra-fast LC 3 Modification Kits 1100 – VWD: 4.6mm RRHT-1100 Series Ultra-fast LC Kit P/N – DAD: 4.6mm RRHT-1100 Series Ultra-fast LC Kit P/N – DAD/MS:2.1mm RRHT-1100 Series Ultra-fast LC Kit P/N Content Filter Capillaries Fittings, Union UV Flow cell RRHT Columns

45 Rapid Resolution HT 1100 Series Modification Kits Converting an 1100 Binary System to Ultra-fast LC Fast LC Modifications for 1110 Binary LC System with DAD/MWD Detector and 4.6mm ID RRHT Columns

46 Summary 1100 Series DAD SL and MWD SL Faster resultswe Higher ResolutionHigher Data Security Higher Peak Capacity  80Hz Data Acquisition for up to 100% resolution gain in ultra-fast LC  High Precision, Linearity and Sensitivity to maintain data quality under ultra-fast LC conditions to comply with regulatory requirements to allow for spectral analysis at trace levels (DAD SL only)  High Instrument, Column and Method Stability enables robust 24x7 operation  Uncompromised Compatibility with Existing Methods Run conventional methods without compromising data quality  Build-in Data Recovery Card provides „data never lost insurance“  RFID Tags for Cells and UV Lamp for unambiguous data traceability  New Low Noise Electronics, New ETC, New Standard Flow Cell for decreased short-term noise and increased practical sensitivity

47 Appendix

48 Overview – Features and Specifications Feature/Spec1100 DAD1100 DAD SL1100 MWD1100 MWD SL Spectral DA Rate10 Hz80 HzN/A Signal DA Rate20 Hz80 Hz20 Hz80 Hz Data Recov. CardNoYesNoYes RFID tagsNoYesNoYes LAN on-boardNoYesNoYes Noise 254nm+/- 10uAU Noise 750nm+/- 10uAU Noise dual WL+/- 10uAU Linearity> 2 AU WL-Range nm WL Accuracy+/- 1nm Dual –LampYes Slit/nm1,2,4,8,16 progr. Note: For uncompromised compatibility from nano-flow to preparative applications all detectors can be ordered with 5 different analytical flow cells (13ul, 5ul, 1.7ul, 500nl, 80nl) and 3 preparative flow cells (3mm, 0.3mm and 0.06mm)

49 Future proof design State-of-the-Art Motorola PowerPC Processor LAN on board USB on board PCMCIA Firewire WLAN Bluetooth Integrated Web-server Module Independent Electronic Core New Electronics platform

50 Peak Width – Response Time – Data Rate and Sensitivity Don‘t use for > 0.15 sec peak width. > 0.15 sec > 0.3 sec > 0.6 sec > 1.2 sec > 3 sec > 6 sec > 12 sec > 24 sec > 51 sec Peak Width = Peak Width at 50% Peak Height Recommended settings in ultra-fast LC with 50% peak width between 0.15 and 0.6 sec For 50% peak width between 0.6 and 1.2 sec Notes: Noise level changes ~ proportional to the square root of the change in data rate. For optimum selectivity and sensitivity the Peak Width should not be chosen smaller than necessary. For 50% peak width between 0.3 and 0.6 seconds Peak Width of > min is recommended, which correspondes to 40Hz data rate. For peaks narrower than 0.3sec at half height, Peak Width of > min (80Hz data rate) should be used. For highest sensitivity in ultra-fast LC the slit can be increased to 8 or 16nm.

51 Linearity - OQ/PV Test on Caffeine Standards min mAU Amount[ng/ul] 0200 Area caffeine, DAD1 A Correlation: Rel. Res%(1): Area = *Amt Correlation:

52 Agilent 1100 Series HT LC/MS System A Scalable and Flexible Solution for Ultra-fast Analysis HT LC System + 1.8um RRHT Columns 2. Capacity Extension 3. Valves 4. Mass-selective Detector 5. Integrated Controller 6. Services and Compliance Products

53 Van Deemter Curves 1.8um Rapid Resolution HT Columns on the 1100 HT System ZORBAX Eclipse XDB-C x 50mm (30mm) 85:15 ACN:Water 1.0  L Octanophenone 0.05 – 5.0 mL/min 20°C 5.0  m 3.5  m 1.8  m 260,740 2mL/min 5.0 mL/min Efficiency gain of 1.8μm versus 5μm columns: 2ml/min 5ml/min HETP (cm/plate) Interstitial linear velocity (u e - cm/sec) Higher speed – Higher resolution – Higher sensitivity ParticleH_min 5μm9.3μm 3.5μm6.0μm 1.8μm3.8μm Note: Efficiency of 1.8μm columns is virtually flow-rate independent.  Up to 2.1x Resolution  Up to 4.4x Speed

54 Ultra-fast Gradient Analysisof 9 Phenones Performance Summary 9 Phenones baseline separated in 29 sec 39 sec 0.8 min (with column regeneration and run time = 0.7min) 1.0 min (without column regeneration and run time = 0.7min) > 2.65 for all peaks 0.50 sec (average) 79 (0.65min gradient) < 0.2% RSD without column regeneration < 1.0% RSD with column regeneration < 0.003% (limit of detection) 7.2 days or 8000 injections of continuous operation with stable performance Analysis speed Analysis time Cycle time Resolution 4  peakwidth Peak capacity RT precision Carry over Robustness min mAU Possible cycle time with CR = 0.8 min Analysis time = 0.65 min = 39 sec Possible cycle time w/o CR = 1.0 min 60°C 4.35ml/min 390bar